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This study aims to investigate the effects and the molecular mechanism of Huangdi Anxiao Capsules(HDAX)-containing serum in protecting the rat adrenal pheochromocytoma(PC12) cells from diabetes-associated cognitive dysfunction induced by high glucose and whether the mechanism is related to the regulation of NOD-like receptor thermal protein domain associated protein 3(NLRP3)-mediated pyroptosis. The PC12 cell model of diabetes-associated cognitive dysfunction induced by high glucose was established and mcc950 was used to inhibit NLRP3. PC12 cells were randomized into control, model, HDAX-containing serum, mcc950, and HDAX-containing serum+mcc950 groups. Methyl thiazolyl tetrazolium(MTT) assay was employed to determine the viability, and Hoechst 33258/PI staining to detect pyroptosis of PC12 cells. Enzyme-linked immunosorbent assay(ELISA) was employed to measure the levels of interleukin-1 beta(IL-1β) and IL-18. Western blot was employed to determine the protein levels of postsynaptic density protein 95(PSD-95), NLRP3, apoptosis-associated speck-like protein containing a CARD(ASC), gasdermin D(GSDMD), GSDMD-N, and cleaved cysteinyl aspartate specific proteinase-1(caspase-1), and RT-PCR to determine the mRNA levels of NLRP3, ASC, GSDMD, and caspase-1. The immunofluorescence assay was adopted to measure the levels and distribution of NLRP3 and GSDMD-N in PC12 cells. Compared with the control group, the model group showed decreased cell proliferation, increased PI positive rate, down-regulated protein level of PSD-95, up-regulated protein levels of NLRP3, ASC, GSDMD-N, GSDMD, and cleaved caspase-1, up-regulated mRNA levels of NLRP3, ASC, GSDMD, and caspase-1, and elevated levels of IL-1β and IL-18. Compared with the model group, HDAX-containing serum, mcc950, and the combination of them improved cell survival rate and morphology, decreased the PI positive rate, down-regulated the protein levels of NLRP3, ASC, GSDMD-N, GSDMD, and cleaved caspase-1 and the mRNA levels of NLRP3, ASC, GSDMD, and caspase-1, and promoted the secretion of IL-1β and IL-18. The findings demonstrated that HDAX-containing serum can inhibit the pyroptosis-mediated by NLRP3 and protect PC12 cells from the cognitive dysfunction induced by high glucose.
Subject(s)
Rats , Animals , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Interleukin-18 , Pyroptosis/physiology , Diabetes Mellitus , Caspases , Glucose , RNA, MessengerABSTRACT
Panax japonicus, which in the Tujia dialect is known as “Baisan Qi” and “Zhujieshen”, is a classic “qi” drug of Tujia ethnomedicine and it has unique effects on disease caused by “qi” stagnation and blood stasis. This paper serves as the basis of further scientific research and development of Panax japonicus. The pharmacology effects of molecular pharmacology were discussed and summarized. P. japonicus plays an important role on several diseases, such as rheumatic arthritis, cancer, cardiovascular agents, and this review provides new insights into P. japonicus as promising agents to substitute ginseng and notoginseng.
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Objective: To study the active triterpenoid saponins of Tujia ethnomedicine Kouziqi (Panax japonicus var. major). The antitumor activity was screened and the relationship between the structure and activity of the compounds was discussed. Methods: The ethanol extract of Kouziqi was isolated by Silica gel, ODS and MCI column chromatograph and purified by preparative HPLC. The structures were elucidated on the basis of spectroscopic analysis and compared with literatures. Using MTT assay to detect the cytotoxicity of 14 compounds in BGC-823, HCT-116, Hela, HepG-2 cells. Results: A total of 14 known compounds were isolated from Tujia ethnomedicine Kouziqi and determined as chikusetsusaponin IVa methyl ester (1), chikusetsusaponin IVa butyl ester (2), chikusetsusaponin IV (3), chikusetsusaponin IVa (4), 28-desglucosylchikusetsusaponin IVa (5), oleanolic acid-3-O-β-D-(6'- methylester)-glucuronopyranoside (6), (24R)-majonoside R1 (7), (24R)-pseudoginsinoside F11 (8), (20S)-notoginsinoside-R2 (9), (20S)-ginsenoside Rg2 (10), ginsenoside Rg1 (11), ginsenoside Re (12), ginsenoside Rd (13) and chikusetsusaponin-V methyl ester (14). Among the 14 compounds, compounds 5 and 6 showed dose-dependent cytotoxicity to BGC-823, HCT-116, Hela and HepG-2 cells. Compound 5 had cytotoxicity in BGC-823 and HCT-116 cells with IC50 values of 9.94 and 14.17 μmol/L, respectively. Compound 6 had the best cytotoxicity in HepG-2 cells with IC50 value of 12.70 μmol/L. Conclusion: Compound 6 is isolated from Kouziqi for the first time and its spectral data were reported. The antineoplastic activity of Tujia ethnomedicine Kouziqi is based on the oleanolic acid-type triterpenoid saponins and related to the substituents of C-28, but the mechanism still needs to be deeply studied.
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Transvaginal NOTES surgery,as an emerging minimally invasive surgical technique,is widely used in gynecology disease.However,there are some difficulties in clinical application. This article discusses the indications and contraindications of transvaginal NOTES,the difficulties and countermeasures in the establishment of the access platform,the exposure problems of the surgical visual field,the complications encountered during the operation and their treatment,and infection prevention.
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AIM To investigate the therapeutic action and mechanism of Shuwei Decoction (Bupleuri Radix,Cyperi Rhizoma,Atractylodis macrocephalae Rhizoma,etc.) on functional dyspepsia (FD) from the perspectives of autophagy and intercellular gap junction pathway through its impact on the expressions of Beclin-1,mTOR,Cx43 protein and mRNA in gastric antrum pyloric smooth muscle cells of FD model rats.METHODS Sixty Wistar rats were randomly divided into control (blank) group,model group,mosapride group (1.37 mg/kg),low,medium and high dose Shuwei Decoction groups (7.67 g/kg,15.34 g/kg,30.68 g/kg,respectively).FD model rats were prepared according to compound etiological modeling method for 21 days.On the first day after modeling,the rats in each group were given the corresponding medicine for intragastric administration once a day for 14 days.Two hours after the last administration,we procured gastric antrum and pyloric tissues from the sacrificed rats.The expressions of Beclin-1,mTOR and Cx43 protein were detected by Western blot,and so were the expressions of Beclin-1,mTOR and Cx43 mRNA by quantitative real-time PCR (RT-qPCR).RESULTS We observed decreased expression of Beclin-1 protein in low,medium and high dose Shuwei Decoction groups (P < 0.01),and an increased expression of Cx43 protein in high dose Shuwei Decoction group (P < 0.05).In medium and high dose Shuwei Decoction groups,decreased expression of Beclin-1 mRNA (P < 0.01),and increased expression of mTOR and Cx43 mRNA (P < 0.01) were detected as well.CONCLUSION Shuwei Decoction can improve FD by suppressing cell autophagy and enhancing the function of gap junction through up-regulating the expression of mTOR mRNA and Cx43 protein and mRNA,and down-regulating the expression of Beclin-1 protein and mRNA.
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<p><b>Background</b>The effectiveness of an anti-incontinence procedure concomitant with prolapse reconstruction for pelvic organ prolapse (POP) in preventing urinary incontinence (UI) after surgery remains controversial. Our study aimed to describe the incidence of pre- and postoperative UI for pelvic reconstructive surgery and evaluate the management of POP associated with UI.</p><p><b>Methods</b>A total of 329 patients who underwent total pelvic reconstruction between June 2009 and February 2015 at a single institution were identified. These patients were divided into two groups (Group A [Prolift reconstruction]: n = 190 and Group B [modified total pelvic reconstruction]: n = 139). Data regarding surgical procedures and patient demographic variables were recorded. Chi-square and Student's t-tests were used for two independent samples.</p><p><b>Results</b>A total of 115 patients presented with UI preoperatively. The average follow-up time was 46.5 months, with 20 patients lost to follow-up (6.1%). The cure rates of stress UI (SUI), urgency UI (UUI), and mixed UI (MUI) were 51% (30/59), 80% (16/20), and 48% (14/29), respectively. The cure rate of UUI after total pelvic reconstruction (80% [16/20]) was higher than that of SUI (50.8% [30/59], χ= 5.219, P = 0.03), and the cure rate of MUI (48%, 14/29) was the lowest. The cure rate of patients with UI symptoms postoperatively was lower than that of those with symptoms preoperatively (9.1% [28/309] vs. 16.2% [50/309], χ= 7.101, P = 0.01). There was no difference in the incidence of UI postoperatively between Groups A and B (P > 0.05). The cure rate of SUI in patients undergoing tension-free vaginal tape-obturator was not higher than that in those who did not undergo the procedure (42.9% [6/14] vs. 53.3% [24/45], χ= 0.469, P = 0.49). There were no differences in the cure rate for POP or UI between these two types of reconstructions (P > 0.05).</p><p><b>Conclusions</b>No correlation between the incidence of UI and POP was identified. The results suggest that UI treatment should be performed after POP surgery for patients with both conditions.</p>
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<p><b>OBJECTIVE</b>To study the expression of DNMT3b gene in myeloma RPMI8226 cells and its biological significance.</p><p><b>METHODS</b>The activity of DNA methyltransferase was detected by ELISA, and the expression of DNMT3b in RPMI8226 cells was analyzed by semi-quantitative RT-PCR and real-time fluorescent quantitative PCR. The proliferation and expression of DNMT3b gene in RPMI8226 cells intervened with capecitabine for 24 hours were detected.</p><p><b>RESULTS</b>The activity of DNMT and expression of DNMT3b in RPMI 8226 cells increased. The proliferation of RPMI8226 cells was inhibited, and the apoptosis occurred in RPMI 8226 cells intervened with capecitabine for 24 hours. The expression level of DNMT3b gene was decreased after being intervened with capecitabine for 24 hours.</p><p><b>CONCLUSION</b>The expression level of DNMT3b in myeloma RPMI 8226 cells increase, and capecitabine can inhibit the proliferation of RPMI 8226 and induce apoptosis by inhibiting the expression of DNMT3b gene. Therefore, DNMT3b is expected to be a new target for myeloma therapy.</p>
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<p><b>OBJECTIVE</b>To investigate the effects of different hemapheresis procedures on the components of hematopoietic stem cells(HSCs) collected from helathy donors.</p><p><b>METHODS</b>twelve donors were underwent stem cell collection from January 2015 to August 2016, and the stem cells were randomly colleted by AutoPBSC procedure of COBE spectra and MNC procedure of the Spectra Optia blood cell separator, the mononuclear cells, CD34cells, granulocytes, lymphocytes and platelets in the collections were compared.</p><p><b>RESULTS</b>The circulating blood volume, the acquisition time and dosage of anticoagulants were not significantly different between two procedures. The volume and the mononuclear cell count collected by AutoPBSC procedure were lower than those by the MNC procedure, while the CD34cell count by AutoPBSC procedure was higher than that by the MNC procedure. More lymphocytes and platelets were collected by AutoPBSC procedure as compared with that by the MNC procedure (P<0.05).</p><p><b>CONCLUSION</b>Compared with MNC procedure of the Spectra Optia blood cell separator, the number of collected stem cells, lymphocytes and platelets are higher by using AutoPBSC procedure of the COBE spectra blood cell separator.</p>
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<p><b>OBJECTIVE</b>To observe morphological changes of enteric nervous system (ENS)-interstitial cells of Cajal (ICC)-smooth muscle cell (SMC) structure injury in deep muscle nerve plexus offunctional dyspepsia (FD) rats, and the repair of Shuwei Decoction (SD) on it, and to explore its effecton FD.</p><p><b>METHODS</b>Totally 72 rats were randomly divided into the control group, the model group, the lowdose SD group, the medium dose SD group, and the high dose SD group, the Mosapride group, 12 ineach group. Rats in the low dose SD group, the medium dose SD group, and the high dose SD group were intragastrically fed with SD at 0.767, 1.534, 3.068 g/mL, respectively. Rats in the Mosapride group were intragastrically fed with Mosapride (1.37 mg/kg). FD rat model with Gan depression Pi deficiency syndrome (GDPDS) was established using complex pathogenic factors. Corresponding liquors were respectively administered to rats in corresponding groups from the 3rd day after modeling. Distilled water(10 mL/kg) was administered to rats in the control group and the model group, once per day for 14 successive days. Rats were sacrificed and small intestine tissues collected for observing ENS-ICC-SMC structure injury using immunofluorescence double labeling, laser scanning confocal microscope, and transmission electron microscope at day 15. Repair of SD on it was also observed.</p><p><b>RESULTS</b>ENS-ICC SMC structure was incomplete, with obvious injury in mutual link of ICC, ICC, SMC, and connecting structure. ENS-ICC-SMC structure was more complete in high, medium, and low dose SD groups, with close link of ICC and SMO. Their connecting structures were in good conditions.</p><p><b>CONCLUSION</b>SD could keep the integrity of ENS-ICC-SMC structure by promoting regeneration and morphology of ICC, thereby, improving gastrointestinal movement disorder and showing therapeutic effect on FD.</p>
Subject(s)
Animals , Rats , Benzamides , Pharmacology , Drugs, Chinese Herbal , Pharmacology , Dyspepsia , Drug Therapy , Enteric Nervous System , Interstitial Cells of Cajal , Morpholines , Pharmacology , Muscle, Smooth , Random AllocationABSTRACT
<p><b>OBJECTIVE</b>To observe the efficacy and adverse reactions of autologous PBSC collection when the autoPBSC procedure and MNC procedure of COBE Spectra cell separator and the MNC procedure of Spectra Optia cell separator were used.</p><p><b>METHODS</b>The autologous perepheral blood hematopoietic stem cells from 41 patients were collected by using autoPBSC procedure and MNC procedure of COBE Spectra blood cell separator and MNC procedure of Spectra Optia blood cell separator. The numbers of MNC and CD34cells collected by 3 collected procedure, the difference of hemoglobin (Hb) drop and platelet decrease, and the adverse reaction of patients were observed.</p><p><b>RESULTS</b>When the whole blood processing and the collection time were basically same among these 3 groups, the MNC counts collected by MNC procedure of COBE Spectra and Spectra Optia were higher than that of AutoPBSC procedure of COBE Spctra, but the CD34cell count was lower than that collected by AutoPBSC procedure (P< 0.05). The final product volume collected by MNC procedure of COBE Spectra and Spectra Optia was bigger than that collected by AutoPBSC procedure. In comprission with MNC procedure of COBE Spectra cell seperator, the CD34count collected by MNC procedure of Spectra Optia Seperator did not show significant difference, but the CD34cell count collected by MNC procedure of Spectra Optia was higher than that collected by MNC procedure of COBE Spectra cell separator (P<0.05). The platelet count and hemoglobin level collected by MNC procedure of Spectra Optia were lower than those before collection. The adverse reactions in the 3 procedures were similar, and the patients could tolerate them.</p><p><b>CONCLUSION</b>The AutoPBSC procedure of COBE Spectra and MNC procedure of Spectra Optia are better than MNC procedure of COBE Spectra for autologous peripheral blood hematopoietic stem cells collection. The loss of blood platelet and hemoglobin after collection is lowest in MNC procedure of Spectra Optia.</p>
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<p><b>BACKGROUND</b>No data on the incidence of pleural effusion (PE) in Chinese patients with pulmonary embolism are available to date. The aim of the current study was to investigate the frequency of PE in a Chinese population of patients with pulmonary embolism.</p><p><b>METHODS</b>This was a retrospective observational single-center study. All data of computed tomography pulmonary angiography (CTPA) performed over 6-year period on adult patients with clinically suspected pulmonary embolism were analyzed.</p><p><b>RESULTS</b>From January 2008 until December 2013, PE was identified in 423 of 3141 patients (13.5%) with clinically suspected pulmonary embolism who underwent CTPA. The incidence of PE in patients with pulmonary embolism (19.9%) was significantly higher than in those without embolism (9.4%) (P < 0.001). Majority of PEs in pulmonary embolism patients were small to moderate and were unilateral. The locations of emboli and the numbers of arteries involved, CT pulmonary obstruction index, and parenchymal abnormalities at CT were not associated with the development of PE.</p><p><b>CONCLUSIONS</b>PEs are present in about one fifth of a Chinese population of patients with pulmonary embolism, which are usually small, unilateral, and unsuitable for diagnostic thoracentesis.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Incidence , Pleural Effusion , Diagnostic Imaging , Epidemiology , Pulmonary Embolism , Diagnostic Imaging , Epidemiology , Radiography , Retrospective StudiesABSTRACT
<p><b>OBJECTIVE</b>To investigate the clinical efficacy of bleomycin, cyclophosphamide, vindesine, Ara-C and dexamethasone(BACOD) for treatment of patients with relapsed/refractory diffuse large B cell lymphoma.</p><p><b>METHODS</b>A total of 56 patients with relapsed/refractory diffuse large B cell lymphoma were divided into control group and treatment group. The control group were received the conventional BACOD treatment, and the treatment group received continuous venoclysis with BACOD. Clinical efficacy was observed and compared.</p><p><b>RESULTS</b>In control group, 6 patients achieved CR, 9 patients achieved PR, and their overall remission rate was 53.6%. In treatment group, 10 patients achieved CR, 12 patients achieved PR, and their overall remission rate was 78.6%. The overall remission rate of treatment group was significantly higher than that of control group (χ2=3.903, P<0.05). The incidence rates of nausea and vomiting, thrombocytopenia, abnormal liver function, fever and granulocytopenia were not significant difference (P>0.05) between the two groups.</p><p><b>CONCLUSION</b>The clinical efficacy of cyclophosphamide, vindesine and dexamethasone for treatment of patients with relapsed/refractory/diffuse large B cell lymphoma has been confirmed to be satisfactory, suggesting that the continuous venoclysis with BACOD can be apptied to the in clinical treatment.</p>
Subject(s)
Humans , Antineoplastic Combined Chemotherapy Protocols , Bleomycin , Cyclophosphamide , Cytarabine , Dexamethasone , Doxorubicin , Lymphoma, Large B-Cell, Diffuse , Recurrence , Treatment Outcome , Vincristine , VindesineABSTRACT
OBJECTIVE@#To observe the preventive and control effect of matrine on transforming growth factor (TGF-β1) and hepatocyte growth factor (HGF) of liver fibrosis tissue in rats.@*METHODS@#A total of 48 SD rats were randomly divided into A, B, C, D groups with 12 in each, group A as the normal control group and groups B, C, D as liver fibrosis models using composite modulus method with carbon tetrachloride (CCL4). Group B was the model group, group C adopted γ-interferon lavage therapy in the second day of modeling, and group D adopted matrine lavage treatment, at 4 and 8 weeks after treatment. Six rats were executed for detection of TGF-β1 and HGF, liver tissue histology and comparison fibrosis degree changes of rat liver tissue between groups.@*RESULTS@#Groups B, C, D showed a more significantly increased TGF-β1 at each time point compared with group A (P<0.05); Group B showed a more significantly increased TGF-β1 than groups C and D at weeks 4 and 8 (P<0.05); group D showed a lowest level of TGF-β1, followed by groups C and B. HGF of group B decreased more significantly than A group at weeks 4 and 8 (P<0.05); HGF of groups C and D was significantly elevated at 4 and 8 weeks than groups A and B (P<0.05), in which the group D showed the highest level of HGF. According to tissue histologic observation, rat liver tissue structure of group A was clear and normal, tissue structure of group B was destroyed with obvious fibrous tissue hyperplasia and fatty change of hepatic cells; groups C and D showed a slighter liver tissue damage, cell necrosis and connective tissue hyperplasia in collect abbacy than group B with a trend of obvious improvement.@*CONCLUSIONS@#Matrine can reduce TGF-β1 expression and enhance the activity of HGF, so as to realize the inhibition effect on liver fibrosis in rats.
Subject(s)
Animals , Male , Rats , Alkaloids , Pharmacology , Gene Expression , Hepatocyte Growth Factor , Genetics , Metabolism , Liver , Chemistry , Metabolism , Pathology , Liver Cirrhosis , Metabolism , Pathology , Protective Agents , Pharmacology , Quinolizines , Pharmacology , RNA, Messenger , Genetics , Metabolism , Rats, Sprague-Dawley , Transforming Growth Factor beta1 , Genetics , MetabolismABSTRACT
Fourteen compoumds were isolated from the ethyl acetate portion of the 95% ethanolic extract of Pleione bulbocodioides by a combination of various chromatographic techniques including silica gel, ODS, macroporous adsorbent resin, Sephadex LH-20, and preparative HPLC, of which ten compoumds were phenanthrenes and dihydrophenanthrenes, two compoumds were bibenzyls, one was lignan and a sterol. Their structures were identified on the basis of spectroscopic data as monbarbatain A(1), 2, 7, 2'-trihy-droxy-4, 4', 7'-trimethoxy-1, 1'- biphenanthrene(2), blestriarene A(3), pleionesin B(4), shanciol H(5), 17-hydroxy-7'-(4'-hy-droxy-3 '-methoxyphenyl)- 4-methoxy-9, 10, 7', 8'-tetrahydrophenanthro[2, 3-b]furan-8'-yl methyl acetate(6), 1-p-hydroxybenzyl-4-methoxy phenanthrene-2, 7-diol(7), 1-p-hydroxybenzyl-4-met-hoxy-9, 10-dihydrophenanthrene-2, 7-diol(8), hircinol(9), coelonin( 10), gigantol(11), batatasin 11 (12), syringaresinol(13) and ergosta4, 6, 8 ( 14) , 22-tetraen-3-one (14). Compounds 1-3, 9, 13 and 14 were isolated from this genus for the first time.
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Drugs, Chinese Herbal , Chemistry , Orchidaceae , Chemistry , Organic ChemicalsABSTRACT
This study was to investigate the chemical constituents from pseudobulbs of Pleione yunnanensis, one of the source of traditional Chinese medicine "Shancigu". The chemical constituents were isolated by various chromatography methods, including silica gel, ODS, Sephadex LH-20, and semi-preparative HPLC. Fourteen compounds were isolated and identified from the EtOAc fraction of 90% ethanol extract, including five dihydrophenanthrenes, four bibenzyls, two triterpenoids, and three phenylacrylic acids. Their structures were identified on the basis of the spectral data as 4, 7-dihydroxy-2-methoxy-9,10-dihydrophenanthrene (1), 4, 7-dihydroxy-1-(p-hydroxybenzyl)-2-methoxy-9,10-dihydrophenanthrene (2), (2,3-trans)-2-(4-hydroxy-3-methoxyphenyl) -3-hydroxymethyl-10-methoxy-2,3,4,5-tetrahydro-phenanthro[2,1-b]furan-7-ol (3), pleionesin B (4), blestriarene A (5), batatasin III (6), 3, 3'-dihydroxy-2-(p-hydroxybenzyl) -5-methoxybibenzyl (7), 3', 5-dihydroxy-2-(p-hydroxybenzyl) -3-methoxybibenzyl (8), 3,3'-dihydroxy-2,6-bis(4-hydroxybenzyl) -5-methoxybibenzyl (9), triphyllol (10), pholidotin (11), (E) -p-hydroxycinnamic acid (12), (E)-ferulic acid (13), and (E)-ferulic acid hexacosyl ester (14). Compounds 5,10-14 were separated from this plant for the first time.
Subject(s)
Drugs, Chinese Herbal , Chemistry , Molecular Structure , Orchidaceae , Chemistry , Spectrometry, Mass, Electrospray IonizationABSTRACT
Larp4b is a member of the LARP family, which can interact with RNA and generally stimulate the translation of mRNA. Abnormal expression of Larp4b can be found in leukemia patients in our previous study. This study was purposed to detect the relative expression of Larp4b mRNA in different subpopulations of mouse hematopoietic cells, to construct lentivirus vector containing shLarp4b targeting mouse gene Larp4b and to explore its effects on mouse Lin(-) cells infected with shLarp4b by lentivirus. SF-LV-shLarP4b-EGFP and control vectors were constructed and two-plasmid lentivirus packing system was used to transfect 293T cells. After 48 h and 72 h, lentivirus SF-LV-shLarp4b-EGFP was harvested and was used to infect Lin(-) cells. After 48 h, EGFP(+) cells was sorted by flow cytometry (FCM). Meanwhile, semi-quantitative real time-PCR, AnnexinV-PE/7-AAD staining, PI staining and colony forming cell assay (CFC) were performed to determine the expression of Larp4b and its effect on the proliferation of hematopoietic progenitor cells. The results showed that Larp4b was highly expressed in myeloid cells. SF-LV-shLarp4b-EGFP was successfully constructed according to the restriction endonuclease digestion assay. RT-PCR confirmed that Larp4b was efficiently knockdown in mouse Lin(-) cells. The low expression of Larp4b did not affect the colony forming number, the apoptosis and cell cycle of Lin(-) cells. It is concluded that knockdown of Larp4b in mouse Lin(-) cells do not contribute to the colony forming ability and the growth of Lin(-) cells in vitro. This useful knockdown system will be used to study in vivo Larp4b in future.
Subject(s)
Animals , Humans , Mice , Autoantigens , Metabolism , Cells, Cultured , Flow Cytometry , Gene Knockdown Techniques , Genetic Vectors , Hematopoietic Stem Cells , Cell Biology , Lentivirus , Genetics , Plasmids , Ribonucleoproteins , Metabolism , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To determine the methods for establishing an in vivo model of long-term hepatitis B virus (HBV) infection in the Chinese tree shrew (Tupaia belangeri chinensis).</p><p><b>METHODS</b>Seventy-seven neonate (1-3 days old) and 49 young adult (2 weeks to 1 year old) tree shrews were inoculated with different HBV sources (chronic hepatitis B (CHB) human patient serum, single or pooled; HBV-infected tree shrew serum, single only; HBV-infected HepG2.2.15 cells' culture medium supernatant; HBV genome-transfected HepG2.2.15 cells' culture medium supernatant) through various routes of injection (subcutaneous, intraperitoneal, and direct liver via abdominal skin; adults also received intravenous and indirect liver via spleen). Serum and liver biopsies were collected from the animals at various time points post-inoculation for detection of HBV markers by fluorescence quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, time-resolved immunofluorescence, Southern blotting, dot blotting, immunohistochemistry, and microscopy.</p><p><b>RESULTS</b>Among the neonatal group of tree shrews, six (7.8%) were confirmed as HBV-infected for more than 72 (up to 228) weeks after inoculation and another seven (9.1%) were suspected of persistent infections. None of the young adult tree shrews developed persistent infection. Inoculation with single-source serum from either CHB humans or tree shrews were responsible for the most cases of infections, and the subcutaneous injection produced more infections than the other inoculation routes. The most reliable methods of determining HBV infection status were detection of serum HBV immunoreactive markers and intrahepatic HBV DNA.</p><p><b>CONCLUSION</b>In order to establish an in vivo model of CHB in the tree shrew, the animals should be inoculated in the neonatal period using subcutaneous injection.</p>
Subject(s)
Animals , Female , Humans , Male , Disease Models, Animal , Hep G2 Cells , Hepatitis B virus , Hepatitis B, Chronic , Virology , TupaiaABSTRACT
<p><b>OBJECTIVE</b>To compare the clinical efficacy differences among acupuncture at special acupoints, nonspecific acupoints and non acupoints in Foot Yangming Meridian for functional dyspepsia (FD) at different time points.</p><p><b>METHODS</b>One hundred and sixteen FD patients were randomly divided into a special acupoints in Foot Yangming Meridian group (group A, n = 36), a non-specific acupoints in Foot Yangming Meridian group (group B, n 39) and a non acupoints group (group C, n = 41). Group A was treated with acupuncture at Chongyang (ST 42), Fenglong (ST 40), Zusanli (ST 36) and Liangqiu (ST 34), and group B was treated with acupuncture at Tiaokou (ST 38), Yinshi (ST 33), Futu (ST 32) and Dubi (ST 35), and the assisted acupoints were setting up at 2 mm apart from selected acupoints in meridian of proximal part. Group C was treated with 4 non acupoints: (1) medial elbow, middle point on the line between olecranon and armpit; (2) middle point on the line between condylus medialis humeri and wrist of elbow-bone; (3) the junction of deltoid and biceps inside the arm; (4) 1-2 cm horizontally away from Zusanli (ST 36), lateral border of shin bone, and the assisted acupoints were setting up at 2 mm away from selected non acupoints along limb longitudinal axis of proximal part. All acupoints and assisted acupoints were connected to the HANS nerve stimulator and they were all treated for 4 courses. The FD Symptom Index (FDI) and MOS 36-Item Short-Form Health Survey (SF-36) after treatment, 1 month and 3 months after treatment were observed and the clinical efficacy was assessed.</p><p><b>RESULTS</b>After treatment, the total effective rate of abdominal fullness after meal, early satiation, upper abdominal pain and upper abdominal burning sensation were 84.8% (28/33), 67.7% (21/31), 76.9% (20/26) and 56.3% (9/16), respectively, in group A, and 45.9% (17/37), 38.7% (12/31), 42.9% (12/28) and 38.5% (5/13), respectively, in group B, and 15.8% (6/38), 18.4% (7/38), 46.1% (12/26) and 16.7% (4/24), respectively, in group C. The total effective rate of abdominal fullness after meal, early satiation and upper abdominal pain in group A were superior to those in group B (all P < 0.05), and the total effective rate of abdominal fullness after meal, early satiation, upper abdominal pain and upper abdominal burning sensation in group A were all superior to those in group C (all P < 0.05), and the total effective rate of abdominal fullness after meal in group B was superior to that in group C (P < 0.05). The score of FDI and SF-36 after treatment, 1 month and 3 months after treatment in all the groups were better than those before treatment (all P < 0.05), and above indices in group A were the most significant (all P < 0.05), and the scores of FDI and SF-36 after treatment in group B were better than those in group C (both P < 0.05).</p><p><b>CONCLUSION</b>All the treatment of acupuncture at special acupoints, non-specific acupoints in Foot Yangming Meridian and non acupoints have therapeutic effect on FD, but acupuncture at special acupoints has better short and long term therapeutic effects, which confirm the existence of acupoints specificity.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Acupuncture Points , Acupuncture Therapy , Digestion , Dyspepsia , Therapeutics , Meridians , Treatment OutcomeABSTRACT
<p><b>BACKGROUND</b>The condition of concomitant upper lobe emphysema and lower lobe fibrosis as identified by computer tomography is known as combined pulmonary fibrosis and emphysema (CPFE). CPFE has distinct clinical characteristics compared with emphysema alone (EA) and idiopathic pulmonary fibrosis (IPF) without emphysema. However, the pulmonary inflammation characteristics of CPFE are not well known, and the differences between CPFE and the other two diseases with regards to pulmonary inflammation need to be explored. The pulmonary inflammatory characteristics were investigated in CPFE patients and compared with EA and IPF.</p><p><b>METHODS</b>Fraction exhaled nitric oxide (Fe,NO) and differential cell counts, the concentrations of monokine induced by interferon gamma (MIG/CXCL9), interferon-inducible protein 10 (IP-10/CXCL10), and interferon-inducible T cell alpha chemoattractant (I-TAC/CXCL11) were measured in induced sputum obtained from subjects with CPFE (n = 22), EA (n = 22), IPF (n = 14), and healthy volunteers (HV, n = 12). In addition, immunohistochemistry was used to quantify the expression of nitric oxide synthases in alveolar macrophages in 23 lung tissues from patients and control subjects.</p><p><b>RESULTS</b>The CPFE group had higher alveolar NO than subjects in the EA and HV groups (P = 0.009, P = 0.001, respectively) but not than the IPF group (P > 0.05). Numbers of sputum eosinophils were significantly elevated in CPFE and IPF groups compared with the HV group (P = 0.001, P = 0.008). In contrast, eosinophil counts in EA group did not differ from those in the HV group. Compared with the EA and HV groups, the CPFE group had a lower concentration of I-TAC/CXCL11 in sputum supernatants (P = 0.003, P = 0.004). Immunoreactivity for inducible nitric oxide synthase (iNOS) was higher in the CPFE group than in the EA group (P = 0.018, P = 0.006, respectively).</p><p><b>CONCLUSIONS</b>The pulmonary inflammation of CPFE group is more similar to IPF group, while the distal airway inflammation is more significant in CPFE and IPF groups than in EA group. Lung eosinophil cell infiltration and high NOS expression in alveolar macrophage might participate in this pathogenesis.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Breath Tests , Chemokines , Immunohistochemistry , Lung , Pathology , Nitric Oxide , Nitric Oxide Synthase Type II , Pneumonia , Pathology , Pulmonary Emphysema , Pathology , Pulmonary Fibrosis , Pathology , Sputum , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the underlying tumor susceptibility mechanisms and reasons for the high risk of cancer in Fanconi anemia (FA).</p><p><b>METHODS</b>Gene Set Enrichment Analysis (GSEA) was performed to compare gene expression profiles between 21 FA patients' bone marrow (BM) mononuclear cell (BMNC) and 11 normal controls in cancer related gene sets from NCBI GEO database, then core enriched genes were identified by further investigation. Through enrichment analyzing biological processes of gene ontology sets and structural genomic gene sets between FA expression profiles and control, more details related with its tumor susceptibility had been revealed.</p><p><b>RESULTS</b>Compared with normal control, gene expression in FA group had significant been enriched in resistance to Bcl-2 inhibitor gene set, fibroblast growth factors signalling pathways, insulin and insulin-like growth factors (IGF) signalling pathways induced cancer genesis gene sets. The high level of D4S234E, SST, FGFs, IGFs, FGFRs and IGFBP expression provided an initiate environment for tumorgenesis and drug resistance. There were significant differences in biogenesis extracellular molecules and cytomembrane structure organizations between FA and control. Genes with promoter regions around transcription start sites containing either motif RRCAGGTGNCV or CCTNTMAGA were enriched and those former genes match annotation for tumorgenic transcription factor 3 (TCF3).</p><p><b>CONCLUSIONS</b>The high tumor susceptibility of FA patients may be closely related with the dramatic changes in cancer related growth factors and hormones environment. This study provides new insights into tumor susceptibility mechanism in FA patients.</p>